Large-scale propagation of Myrobolan (Prunus cerasifera) in RITA® bioreactors and ISSR-based assessment of genetic conformity

An efficient protocol for the micropropagation of Prunus cerasifera using temporary immersion bioreactors was described. One-node cuttings excised from field-grown plants were successfully disinfected using 12% sodium hypochlorite for 10 min. Immersion duration, frequency and medium composition in Bioreactors type RITA® were evaluated. A high multiplication rate was obtained using RITA® bioreactors containing Murashige and Skoog's medium (MS) supplemented with 1 mg L−1 N6-benzyladenine (BAP) and 1 mg L−1 indole-3-butyric acid (IBA) with 20 min/12 h as immersion time. RITA®-derived shoots were found to be more vigorous than those regenerated using the standard procedure. Higher levels of photosynthetic pigments were observed in shoots cultured in bioreactors, during the proliferation and rooting stages, which proved a certain degree of photoautotrophy of RITA®-derived vitroplants. A high proportion of shoots was successfully rooted in an MS liquid medium supplemented with 1 g L−1 activated charcoal in RITA® bioreactors. During plant acclimatization, survival rates exceeding 80% were recorded. The genetic fidelity of micropropagated plants was investigated using inter simple sequence repeat (ISSR) markers. Results proved the generation of homogenous amplification profiles and thus supported the clonal fidelity of regenerated vitroplants.